Methods for three dimensional reconstruction and determining the quality of an embryo
a three-dimensional reconstruction and embryo technology, applied in the field of reproductive medicine, can solve problems such as failure of implantation, complete disappointment, and limitations to predict success
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example 1
[0068]Embryoscan.
[0069]The present invention relates to the methods for three dimensional (3D) reconstruction and 3D printing of human oocytes and embryos. More precisely, it pertains to the use of 3D technologies as new and innovative tools to improve oocyte and embryo morphology assessment during their preimplantation development in vitro.
[0070]The present invention is based on 3D reconstruction and 3D printing of human oocytes and embryos. These 3D technologies can be used for assessing embryo morphology and improving embryo selection in order to increase ART success. Moreover, 3D technologies represent an informative tool for training, updating of embryologist staff and for patients undergoing ART procedure.[0071]This invention comprises the steps consisting of:[0072]i) Providing serially image sections of human oocyte and embryo from standard optical microscopy or light sheet microscopy (for example SPIM) or Optical Coherence Tomography (OCT) or Magnetic Resonance Micro Imaging...
example 2
[0094]Example of a microscope using like sheet based on SPIM. An optical device (1) produces a light sheet (2), illumining a single plane of the object (4). Two objectives (3), placed at 90° relative to the light sheet beam, are each connected to CCD cameras. A computer receives the images from the two cameras and pilots automatically the displacements of the motorized xyz stage (5) to make stacks of pictures of the different planes of the embryo alternatively illuminated by the SPIM. The pictures are finally sent into a visualization software: it mixes the pictures of the two cameras and shows the result in Multiplanar Reconstruction (MPR) and 3D (VR) (FIG. 4).
example 3
[0095]In Vitro Fertilization, Embryo Quality Classification and IVF Outcomes.
[0096]Ovulation is induced by a single injection of 250 μg of human chorionic gonadotropin (OVITRELLE®; Merck Serono). Oocyte retrieval is performed by transvaginal ultrasound-guided aspiration 36 hours after the injection and each pre-ovulatory follicle is aspirated individually without flushing.
[0097]Cumulus-oocyte complexes are isolated for conventional IVF or ICSI procedures. Before microinjection for ICSI, oocyte maturity is assessed after denudation. Oocytes are individually cultured in a 30 μl microdroplet of culture medium (VITROLIFE®) under oil at 37° C. in 6% CO2 and humid atmosphere. Normal fertilization is confirmed by the presence of two pronuclei and two polar bodies 18 to 20 hours after microinjection or insemination. Early cleavage is observed at 25 or 27 hours after microinjection or insemination, respectively.
[0098]Three days after oocyte retrieval, embryo quality is graded from 1 to 4, ac...
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